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Method for preventing and treating crop hemiptera injurious insects by using Bt proteins

A technology for crops and hemiptera, applied in the field of using Bt protein to prevent and control crop hemiptera pests, can solve the problem of no anti-hemiptera insect activity, etc., and achieve the effect of reducing agricultural production costs and reducing yield loss

Active Publication Date: 2018-03-23
HANGZHOU RUIFENG BIOTECH LIMITED
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Cry30 protein is a class of Bt insecticidal crystal proteins. So far, 13 proteins such as Cry30Aa1 and Cry30Ba1 have been discovered and named. Among them, except for Cry30Fa1, there are no reports of anti-hemipteran insect activity.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Embodiment 1, construct a vector that Agrobacterium transforms rice to express insect-resistant gene

[0018] According to rice codon preference, three insect-resistant protein coding genes, cry30Aa1, cry30Db1 and cry30Ea2, were designed (the amino acid sequence of the protein encoded by the gene is shown in SEQ ID NO.1-3, and its 5' end was designed with a BamHI site, 3 The SacI site was designed at the 'end, and the gene fragment can be obtained by double digestion with BamHI-SacI), which was artificially synthesized by Shanghai Sangong. For rice, the rice actin 1 promoter (pOsActin1, Genbank: NC_008398) was selected to initiate the expression of cry30Aa1, cry30Db1, cry30Ea2 genes, and its 5' end was designed with a HindIII site, and its 3' end was designed with a BamHI site. HindIII-BamHI fragment; maize phosphoenolpyruvate carboxylase terminator (PEPC-ter, Genebank: X15239) was selected as the terminator of the gene, the 5' end was designed with a SacI site, and the...

Embodiment 2

[0020] Embodiment 2, the acquisition of transgenic rice

[0021] The method of obtaining transgenic rice is to adopt the existing technology (Lu Xiongbin, Gong Zuxun, 1998 Life Science 10: 125-131; Liu Fan et al., 2003 Molecular Plant Breeding 1: 108-115). Ripe and plump rice seeds were selected for hulling, and callus was induced as transformation material. Get the Agrobacterium containing the target gene (pCambia1300-pOsActin1-cry30Aa1, pCambia1300-pOsActin1-cry30Db1, pCambia1300-pOsActin1-cry30Ea2 prepared in Example 1) to draw a plate, and pick a single clone colony to inoculate and prepare to transform Agrobacterium. Put the rice callus to be transformed into OD 600 In the Agrobacterium liquid (containing acetosyringone) between 0.3-0.4, let the Agrobacterium bind to the surface of the callus, then transfer the callus to the co-culture medium, and co-culture for 2-3 days. Wash the transformed callus with sterile water, transfer to the selection medium containing antibio...

Embodiment 3

[0022] Embodiment 3, the mensuration of insect resistance ability of transgenic crops

[0023] Utilize brown planthopper and white-backed planthopper to measure the insect-resistant activity of the 10 transgenic rice lines obtained in Example 2, the assay method is: the newly hatched nymphs of brown planthopper and white-backed planthopper are connected to transgenic rice and non-transgenic rice in the same growth period For rice, each strain was placed in a separate insect cage, cultured in a culture room with a constant temperature of 25°C and a constant humidity of 85% for 5 to 7 days, and recorded data. Eight of the 10 transgenic lines tested were not harmed by insects, and all the insects died; all insects on the non-transgenic control group survived, and the rice was severely damaged. The mortality rate of brown planthoppers is shown in Table 1.

[0024]Table 1 Mortality of BPH bioassay on transgenic rice

[0025] Genetically modified rice containing insect-re...

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PUM

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Abstract

The invention discloses a method for preventing and treating crop hemiptera injurious insects by using Bt proteins; the method comprises that the Bt insect-resistant proteins Cry30Aa1, Cry30Db1 and Cry30Ea2 are expressed in crops. Through verification by laboratory studies, the Cry30Aa1, Cry30Db1 and Cry30Ea2 proteins are found to have the activity to resist hemiptera insects, especially delphacidae insects. Encoding genes of the Bt insect-resistant proteins are transferred into the crops, so that transgenic plants (such as paddy rice and the like) can be allowed to obtain the ability to resist the hemiptera insects, especially the delphacidae insects, production loss in agricultural production is reduced, and the cost of agricultural production is reduced.

Description

(1) Technical field [0001] The invention relates to a method for using Bt protein to prevent and control crop hemiptera pests, in particular to a method for expressing an anti-insect protein in rice to enable rice to gain resistance to planthopper insects, and to prevent and control rice planthopper pests. (2) Background technology [0002] Pests cause an annual loss of about 8 billion US dollars to global agricultural production. The control of pests currently mainly relies on the use of chemical pesticides, but the residues of pesticides will have adverse effects on human health and the environment. The use of biotechnology to cultivate transgenic insect-resistant crops containing insect-resistant genes can greatly reduce the use of chemical pesticides and effectively protect crops from pests. Currently, commercialized transgenic insect-resistant corn, soybeans, and cotton Planting has been promoted on a large scale. [0003] In the field, there are many types of Hemipter...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/84C07K14/325A01H5/00A01H6/46
CPCC07K14/325C12N15/8286
Inventor 许超沈志成
Owner HANGZHOU RUIFENG BIOTECH LIMITED